Journal of Cancer Research and Therapeutics

ORIGINAL ARTICLE
Year
: 2022  |  Volume : 18  |  Issue : 2  |  Page : 370--377

SOX7 modulates the progression of hepatoblastoma through the regulation of Wnt/β-catenin signaling pathway


Juntao Ge1, Qipeng Zheng2, Hongyi Qu3, Zuohui Zhao3, Yue Xu3, Hao Wang3, Huayu Gao3, Jianghua Zhan4,  
1 Graduate School, Tianjin Medical University, Tianjin; Department of Pediatrics, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Shandong Engineering and Technology Research Center for Pediatric Drug Development, Jinan; Department of General Surgery, Tianjin Children's Hospital, Tianjin, China
2 Graduate School, Tianjin Medical University; Department of General Surgery, Tianjin Children's Hospital, Tianjin, China
3 Department of Pediatrics, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Shandong Engineering and Technology Research Center for Pediatric Drug Development, Jinan, China
4 Department of General Surgery, Tianjin Children's Hospital, Tianjin, China

Correspondence Address:
Jianghua Zhan
Department of General Surgery, Tianjin Children's Hospital, LongYan Road 238, Beichen District, Tianjin, 300134
China

Abstract

Aims: Hepatoblastoma (HB) was reported as the frequently diagnosed primary hepatic malignant tumor among children. No reports have shown the function of SOX7 and its relationship with the Wnt/β-catenin pathway in HB. Materials and Methods: SOX7 and factors related to Wnt/β-catenin pathway were detected using reverse transcription–quantitative polymerase chain reaction (RT-PCR) and Western blotting. MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium and flow cytometry were used to detect HB cell proliferation and apoptosis. The transwell assay uses cell invasion. Results: In this study, RT-PCR, Western blotting, and immunohistochemistry results indicated that the expression of SOX7 was significantly reduced in HB tissues compared with adjacent noncancerous tissues, while the β-catenin was significantly increased in HB tissues compared with adjacent noncancerous tissues. There were significant differences in the PRETEXT stage and tumor metastasis between patients with low expression and high expression of SOX7. Moreover, it was found that the overexpression of SOX7 and inhibiting Wnt/β-catenin pathway significantly reduced the cell proliferation and invasion, while the cell apoptosis was significantly increased compared with the control group. Conclusions: This study shows that SOX7 was downexpressed in HB tumor tissues. Moreover, ex vivo experiments indicated that SOX7 was related to β-catenin and regulated the progression of HB cells.



How to cite this article:
Ge J, Zheng Q, Qu H, Zhao Z, Xu Y, Wang H, Gao H, Zhan J. SOX7 modulates the progression of hepatoblastoma through the regulation of Wnt/β-catenin signaling pathway.J Can Res Ther 2022;18:370-377


How to cite this URL:
Ge J, Zheng Q, Qu H, Zhao Z, Xu Y, Wang H, Gao H, Zhan J. SOX7 modulates the progression of hepatoblastoma through the regulation of Wnt/β-catenin signaling pathway. J Can Res Ther [serial online] 2022 [cited 2022 Aug 19 ];18:370-377
Available from: https://www.cancerjournal.net/text.asp?2022/18/2/370/345530


Full Text



 Introduction



Hepatoblastoma (HB) was reported as the frequently diagnosed primary hepatic malignant tumor among children. The number of patients with HB accounts for about 0.8%–2.0% of all malignancies in children and almost all malignant hepatic tumors.[1],[2] Currently, although most patients underwent routinely combinatorial chemotherapy and radiotherapy, surgical resection of the HB tumors combined with other options, such as immunotherapy,[3] was the major treatment for patients with HB. Even after surgery, HB patients still have a high possibility of tumor recurrence or distant metastasis.[4],[5] The overall survival of patients with HB has no significant improvement. The primary reason for that is that most children with HB are diagnosed during the late stage, such as PRETEXT (Pre Treatment of extent of disease staging system) III or IV.[6],[7] Lacking effective means of early diagnosis and intervention is a pivotal reason that contributed to the relatively worse prognosis for patients with HB. Novel biomarkers are required for patients with HB to develop diagnostic and prognostic prediction.

SOX7 is a member of the SOX (SRY-related HMG-box) family of transcription factors involved in various physiological processes, including cell survival, proliferation, and cell death.[8],[9],[10] Recently, SOX7 has drawn more attention since it is related to different malignant cancers, including breast cancer, ovarian cancer, liver, and pancreatic cancer,[11],[12],[13],[14] but the mechanisms are largely unclear. The human SOX7 gene is located at the chromosomal region of 8p23.1 and is about 7.7 kbps in length.[12],[15] Based on previous reports, on the one hand, genome-wide promoter analysis indicated that SOX7 was associated with numerous transcription factors, including E2F1, E2F4, and c-MYC., which were closely related to tumorigenesis.[16],[17] Alternatively, a few studies have reported that SOX7 overexpression was related to a favorable prognosis and SOX7 functioned as a tumor suppressor gene. Moreover, SOX7 was reported to associate with the aberrant activation of the Wnt/β-catenin pathway.[16],[18],[19] The Wnt/β-catenin pathway used Wnt ligand and correlated receptors to initiate intracellular signaling through nuclear translocation of β-catenin.[20],[21],[22],[23],[24],[25] The Wnt/β-catenin pathway had been repeatedly reported in patients with HB, while no study focused on the SOX family.[26],[27] SOX7 has a significant role in influencing the factors related to the Wnt/β-catenin signaling pathway, especially the β-catenin, in the cacogenesis. Nevertheless, no reports have demonstrated the function of SOX7 and its interaction with the β-catenin in HB.

In this study, an analysis of the expression of SOX7 and β-catenin in patient samples with HB was conducted and the potential interaction between SOX7 and β-catenin in cell proliferation and survival was further explored.

 Materials and Methods



Samples from children with hepatoblastoma

Twelve paired HB tissues accompanied by adjacent noncancerous tissues were obtained and analyzed from patients with HB who received hepatic surgical resection at the hospital. Patients did not receive any treatment before tumor resection. There are five female and seven male patients. The stages of patients with HB were based on the pretreatment extension stage (PRETEXT) system. This PRETEXT system was identified as the essential and significant assessment method for risk stratification and prognostic prediction for patients with HB. It described the potential extent of HB tumor and the location of all tumors in the liver. Moreover, it included many annotation factors to involve the related tumor characters, such as vascular involvement and lymph invasions, portal vein or hepatic vein invasion or destruction, distant metastasis, the number of tumors, and tumor rupture.[28],[29],[30] Two independent pathologists conducted the histology and pathology evaluation of the tumors from all children. In this study, the adjacent noncancerous tissues were based on the literature and defined as sampled ≥1-cm away from the primary tumor and the tumor margin also identified by two independent pathologists. This study was approved by the local ethics committee of the hospital.

Cell culture and transfection

HB cell lines (HepG2 and Huh6) were purchased from the American Type Culture Collection (ATCC; Rockville, Md, USA). The cell culture methods and selecting the appropriate medium and other materials were followed with the manufacturer's instructions and previously published articles.[31],[32],[33] The lentivirus packaging of SOX7 mimics, NC mimics, or β-catenin inhibitors were purchased from the company of RiboBio, and the transfection of these lentiviruses with HepG2 and Huh6 followed the reagent instructions supplied by the company.

Extract the total RNA

The Recover All™ Total Nucleic Acid Isolation Kits were used to conduct total RNA extraction. All noncancerous samples, tumor samples, and samples from transfection of these lentiviruses with HepG2 and Huh6 were extracted. The maximum absorption wavelength of nucleic acid is 260 nm, which can be used to calculate the concentration of nucleic acid samples. We used Recover All™, the Total Nucleic Acid Isolation Kit to extracts total RNA from all HB tissues and adjacent noncancerous tissue samples. The maximum absorption wavelength of nucleic acid is 260 nm, which can be used to calculate the concentration of nucleic acid samples. The purity of nucleic acid can be estimated by measuring the ratio of optical density (OD) values at 260 nm and 280 nm. Our experimental result indicated that the extracted RNA content is higher. 260/280 analysis shows that the RNA is not degraded, the purity is higher, and there is no DNA pollution.

MTT assays

MTT assays were used to detect the cell proliferation for HepG2 and Huh6 transfected with various lentiviruses and divided into different groups. After being transfected with SOX7 mimics, NC mimics, or β-catenin inhibitor, the HepG2 and Huh6 cells were harvested and the plantation of cells into a 96-well plate was performed. After that, the OD 490 was detected and analyzed using the related software in the microplate reader (Bio-Rad, Hercules, CA, USA).

Immunohistochemistry

Using the tissue slice digital scanner or imaging system to obtain the scanned files or images on the immunohistochemical slices, Seville image analysis system was used to automatically read the tissue measurement area, analyze, and calculate the number of weak, medium, and strong positive cells in the measurement area (negative no Coloring, 0 points; weak positive light yellow, 1 point; medium positive brown, 2 points; and strong positive tan, 3 points), the total cell number, positive cumulative OD IOD value, positive pixel area, and tissue area mm2. The following results were calculated separately to reflect the degree of positiveness. The following indicators can be selected to evaluate the intensity of positive cells according to the slice conditions.

Histochemistry score (H-score) was used to identify the expression of SOX7 and β-catenin. It is a histological scoring method for immunohistochemistry (IHC), which converts the number of positives in each section and its staining intensity into corresponding values to achieve half of the staining of tissue quantitative purposes. H-Score (H-SCORE=Σ(pi × i) = (percentage of weak intensity cells × 1) + (percentage of moderate-intensity cells × 2) + (percentage of strong intensity cells × 3), where i means positive cell grade division: negative without staining, score 0; weak positive light yellow, score 1; medium positive brown, score 2; strong positive tan, score 3. pi represents the percentage of positive cells). H-score is a value between 0 and 300; the larger the value, the stronger the comprehensive positive strength.

Flow cytometry for apoptosis detection

The treated cells were digested and collected and 50,000–100,000 cells (1000 R/min, 5 min) were centrifuged. The supernatant was abandoned and 195-μ L annexin V-FITC binding solution was joined, the cells were gently resuspended. Subsequently, 5-μ L annexin V-FITC reagent was added, gently mixed well, and placed in the dark at room temperature for ten minutes. Then, propidium iodide staining solution 10 μL was added. It was mixed well and kept away from light at room temperature for 10 min. Then, by gently adding 200-μ L annexin V-FITC binding solution, resuspended cells, the cell suspension was then detected using flow cytometry.

Reverse transcription–quantitative polymerase chain reaction

Total RNA was extracted from HB tissues and adjacent noncancerous tissues, and HepG2 and Huh6 cells, followed by the manufacturer's instructions. The following primer pairs were used to perform quantitative polymerase chain reaction (qPCR): β-catenin forward, 5'-GCTGAAGGTGCTATCTGTCTGC-3' and reverse, 5'-CCTTCCATCCCTTCCTGTTTAG-3'; SOX7 forward, 5'-CGAGCTGTCGGATGGACAATC-3' and reverse, 5'-CCACGACTTTCCCAGCATCTT-3'; ACTIN forward, 5'-CACCCAGCACAATGAAGATCAAGAT-3' and reverse, and 5'-CCAGTTTTTAAATCCTGAGTCAAGC-3'.

Western blotting analysis

Tumor tissue was collected from patients and adjacent noncancerous tissues in radioimmunoprecipitation assay; total protein was buffered and extracted. The protein concentration was calculated using the bicinchoninic acid protein determination kit (Thermo Fisher Scientific, Waltham, MA, USA). Then, the protein was electrophoresed and transferred to a polyvinylidene fluoride membrane (Roche, Basel, Switzerland) and sealed with 5% skim milk. Buffered Saline Tween (tbst) was placed at room temperature for 2 h and incubated with a specific primary antibody at 4°C overnight. The following primary antibodies were used: SOX7 (Abcam, 1:200), β-catenin (Abcam, 1:400 and Thermo Scientific, 1:200), E-cad (BD, 1:300), and bcl-2 (Abcam, ab92574, 1:400). ImageJ software was used to calculate the gray value of the Western blotting result band and to conduct statistical analysis. The gray value of the target protein is divided by the gray value of the internal reference to correct the error. The result represents the relative content of the target protein of a sample.

Statistical methods

All data were indicated as the mean ± standard error of mean. Statistical analysis was conducted using GraphPad Prism software. Continuous variables were expressed as mean ± standard deviation and compared using a two-tailed unpaired Student's t-test; categorical variables were compared using Chi-Squared 2 or Fisher analysis.

 Results



Comparison of SOX7 and β-catenin expression in adjacent noncancerous and tumor tissues in Hepatoblastoma patients

The qPCR assay results indicated that the expression of SOX7 was significantly reduced in HB tissues compared with adjacent noncancerous tissues, while the β-catenin was significantly increased in HB tissues compared with adjacent noncancerous tissues [Figure 1]a. Furthermore, IHC and Western blotting were conducted, and it was found that the expression of SOX7 was significantly reduced in HB tissues compared with adjacent noncancerous tissues, while the β-catenin was significantly increased in HB tissues compared with adjacent noncancerous tissues [Figure 1]b and [Figure 1]c. This finding suggested that SOX7 and β-catenin were related and involved in the development of HB. The expression levels of SOX7 and β-catenin were significantly negatively correlated, which was consistent in the reverse transcription–PCR (RT-PCR), IHC, and Western blotting results.{Figure 1}

Cell transfection was then found and the relative expression of SOX7 and β-catenin using RT-PCR was detected. It was explored that the expression of SOX7 and β-catenin in different groups showed significant differences, proving the transfection rate of SOX7 and β-catenin.

After detection using RT-PCR and IHC, expression of SOX7 and b-catenin-related markers were confirmed by Western blotting in all 12 patients. It was found that the protein levels of SOX7 in the tumor tissues were significantly decreased compared with those in the adjacent noncancerous tissues (P < 0.05), while β-catenin in the tumor tissues was significantly increased compared with those in the adjacent noncancerous tissues.

SOX7 expression was associated with clinical characteristics in patients with hepatoblastoma

In this study, the expression of SOX7 in 12 patients was detected. The baseline characteristics of patients during diagnosis are summarized in [Table 1]. Overall, these patients had a median age of 1.8 years. The gender distribution was roughly equal (male: female: 58.3%:41.7%). The PRETEXT status of most patients was PRETEXT III (six patients, 50%), and PRETEXT 3 (25%). Most patients had solitary tumors (85.7%), with no metastases (75%). Most patients had epithelial histological subtypes (58.3%). After being detected by IHC and Western blotting, it was found that seven patients had low expression of SOX7 and five patients had high expression of SOX7. Particularly, there were significant differences in the PRETEXT stage and tumor metastasis between patients with low expression and high expression of SOX7 [P < 0.05 [Figure 2]].{Table 1}{Figure 2}

SOX7 is closely related to the β-catenin expression in HEPG2 and HUH6 cell lines

Gene overexpression is the process that leads to the abundant target protein expression subsequently. In this study, the SOX7 has been overexpressed through the construction of an expression vector and then transfected with HEPG2 and HUH6 cell lines. It was found that SOX7 was significantly increased and β-catenin was significantly decreased in the overexpression SOX7 group [Figure 3]a and [Figure 3]b. In addition, β-catenin was downexpressed through shRNA in HEPG2 and HUH6 cell lines. It was found that compared with the control group, SOX7 was significantly increased and β-catenin was significantly reduced in the overexpression SOX7 group [Figure 3]c and [Figure 3]d.{Figure 3}

SOX7 regulates the β-catenin expression to inhibit proliferation and invasion and to promote apoptosis of HepG2 and Huh6 cells

HepG2 and Huh6 cells were successfully transfected with lentivirus of SOX7 mimic, NC mimic, β-catenin inhibitor, and NC inhibitor. The MTT assay, transwell assay, and flow cytometry were performed. The results showed that overexpression of SOX7 significantly decreased cell proliferation and cell invasion. Knockdown β-catenin significantly inhibited cell proliferation and cell invasion. The apoptosis results were opposite in these two groups (P < 0.05) [Figure 4] and [Figure 5].{Figure 4}{Figure 5}

 Discussion



Among children, HB is reported as the most commonly diagnosed primary malignant live tumor in children. Although most patients underwent routinely combinatorial chemotherapy and radiotherapy, patients with HB have a slight improvement in prognosis.

The Wnt/β-catenin signaling pathway has been reported as a significant pathway related to cancer genesis in various cancers.[34],[35],[36],[37],[38] Nevertheless, activation of this pathway can cause oncogenic or tumor-suppressive effects of different cancer entities. Several studies have revealed different effects of the Wnt/β-catenin signaling pathway. In HB, there is much conflicting evidence on the role of Wnt/β-catenin signaling, and the activation and inhibition of this pathway are considered a potential treatment strategy.[39],[40] In this study, in vitro and in vivo experiments were conducted. RT-PCR, Western blotting, and IHC results showed that the expression of SOX7 was significantly reduced in HB tissues compared with adjacent noncancerous tissues, while the β-catenin was significantly increased in HB tissues compared with adjacent noncancerous tissues. Concerning the clinical relationship of the expression of SOX7 and β-catenin, the clinical characteristics of patients with HB, there were significant differences in the PRETEXT stage and tumor metastasis between patients with low expression and high expression of SOX7.

SOX7 protein contains a β-catenin interaction region, which can suppress Wnt signaling.[15],[41] This is the theoretical basis of our study to perform in patients with HB. It was discovered that SOX7 is frequently inactivated in HB cell lines, and it was hypothesized that it might act as a tumor suppressor. Suppression of the Wnt/β-catenin signaling pathway by SOX7 has now been observed in many other cancer types, and the down-regulation of SOX7 expression is also correlated with the progression of patients with various cancers. In this study, the expression of SOX7 and factors in the Wnt/β-catenin signaling pathway were detected in patients with HB. However, progression analysis was not performed since there was a limited sample size.

By the way, this study has some limitations, and future studies should be conducted. First, this was a retrospective study, and more patients should be recruited to verify these results. Second, the association between SOX7 and the potential possibilities of β-catenin suppressing HB was not clearly explained. Therefore, further research is needed to explore the potential mechanism for this significant phenomenon.

 Conclusions



Our study demonstrates that SOX7 was downexpressed in HB tumor tissues, and the overexpression of SOX7 related to the early stage of patients with HB. Moreover, ex vivo experiments demonstrated that SOX7 was related to β-catenin and regulated the progression of HB cells.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

References

1Otte JB, Pritchard J, Aronson DC, Brown J, Czauderna P, Maibach R, et al. Liver transplantation for hepatoblastoma: Results from the international society of pediatric oncology (SIOP) study SIOPEL-1 and review of the world experience. Pediatr Blood Cancer 2004;42:74-83.
2Tiao GM, Bobey N, Allen S, Nieves N, Alonso M, Bucuvalas J, et al. The current management of hepatoblastoma: A combination of chemotherapy, conventional resection, and liver transplantation. J Pediatr 2005;146:204-11.
3Horton JD, Lee S, Brown SR, Bader J, Meier DE. Survival trends in children with hepatoblastoma. Pediatr Surg Int 2009;25:407-12.
4Khaderi S, Guiteau J, Cotton RT, O'Mahony C, Rana A, Goss JA. Role of liver transplantation in the management of hepatoblastoma in the pediatric population. World J Transplant 2014;4:294-8.
5Yang W, Chen Y, Huang Y, Wang H. Analysis of factors related to recurrence of paediatric hepatoblastoma – A single centre retrospective study. BMC Pediatr 2019;19:485.
6Maibach R, Roebuck D, Brugieres L, Capra M, Brock P, Dall'Igna P, et al. Prognostic stratification for children with hepatoblastoma: The SIOPEL experience. Eur J Cancer 2012;48:1543-9.
7Qiao GL, Li L, Cheng W, Ge J, Zhang Z, Wei Y. Predictors of survival after resection of children with hepatoblastoma: A single Asian center experience. Eur J Surg Oncol 2014;40:1533-9.
8Otsubo T, Akiyama Y, Yanagihara K, Yuasa Y. SOX2 is frequently downregulated in gastric cancers and inhibits cell growth through cell-cycle arrest and apoptosis. Br J Cancer 2008;98:824-31.
9Thein DC, Thalhammer JM, Hartwig AC, Crenshaw EB 3rd, Lefebvre V, Wegner M, et al. The closely related transcription factors Sox4 and Sox11 function as survival factors during spinal cord development. J Neurochem 2010;115:131-41.
10Stevanovic M, Drakulic D, Lazic A, Ninkovic DS, Schwirtlich M, Mojsin M. SOX transcription factors as important regulators of neuronal and glial differentiation during nervous system development and adult neurogenesis. Front Mol Neurosci 2021;14:654031.
11Liu H, Yan ZQ, Li B, Yin SY, Sun Q, Kou JJ, et al. Reduced expression of SOX7 in ovarian cancer: A novel tumor suppressor through the Wnt/β-catenin signaling pathway. J Ovarian Res 2014;7:87.
12Stovall DB, Cao P, Sui G. SOX7: From a developmental regulator to an emerging tumor suppressor. Histol Histopathol 2014;29:439-45.
13Wang J, Zhang S, Wu J, Lu Z, Yang J, Wu H, et al. Clinical significance and prognostic value of SOX7 expression in liver and pancreatic carcinoma. Mol Med Rep 2017;16:499-506.
14Qin CX, Yang XQ, Zhan ZY. Connection between SOX7 expression and breast cancer prognosis. Med Sci Monit 2020;26:e921510.
15Peng W, Guo L, Tang R, Liu X, Jin R, Dong JT, et al. Sox7 negatively regulates prostate-specific membrane antigen (PSMA) expression through PSMA-enhancer. Prostate 2019;79:370-8.
16Wat JJ, Wat MJ. Sox7 in vascular development: Review, insights and potential mechanisms. Int J Dev Biol 2014;58:1-8.
17Yang Y, Andersson P, Hosaka K, Zhang Y, Cao R, Iwamoto H, et al. The PDGF-BB-SOX7 axis-modulated IL-33 in pericytes and stromal cells promotes metastasis through tumour-associated macrophages. Nat Commun 2016;7:11385.
18Chan DW, Mak CS, Leung TH, Chan KK, Ngan HY. Down-regulation of Sox7 is associated with aberrant activation of Wnt/b-catenin signaling in endometrial cancer. Oncotarget 2012;3:1546-56.
19Bai QL, Hu CW, Wang XR, Shang JX, Yin GF. MiR-616 promotes proliferation and inhibits apoptosis in glioma cells by suppressing expression of SOX7 via the Wnt signaling pathway. Eur Rev Med Pharmacol Sci 2017;21:5630-7.
20Guo Y, Xiao L, Sun L, Liu F. Wnt/beta-catenin signaling: A promising new target for fibrosis diseases. Physiol Res 2012;61:337-46.
21Arend RC, Londoño-Joshi AI, Straughn JM Jr., Buchsbaum DJ. The Wnt/β-catenin pathway in ovarian cancer: A review. Gynecol Oncol 2013;131:772-9.
22Krishnamurthy N, Kurzrock R. Targeting the Wnt/beta-catenin pathway in cancer: Update on effectors and inhibitors. Cancer Treat Rev 2018;62:50-60.
23Cai K, Jiang L, Wang J, Zhang H, Wang X, Cheng D, et al. Downregulation of β-catenin decreases the tumorigenicity, but promotes epithelial-mesenchymal transition in breast cancer cells. J Cancer Res Ther 2014;10:1063-70.
24Fernando G, Paul F, Laura J, Alejandra AM, Gabriela M, Alberto PL. Is the Wnt/β catenin signalling pathway activated in seminoma?: An immunohistochemical study. J Cancer Res Ther 2016;12:1075-9.
25Salehi Z, Akrami H. Target genes prediction and functional analysis of microRNAs differentially expressed in gastric cancer stem cells MKN-45. J Cancer Res Ther 2017;13:477-83.
26Matsumoto S, Yamamichi T, Shinzawa K, Kasahara Y, Nojima S, Kodama T, et al. GREB1 induced by Wnt signaling promotes development of hepatoblastoma by suppressing TGFβ signaling. Nat Commun 2019;10:3882.
27Sha YL, Liu S, Yan WW, Dong B. Wnt/β-catenin signaling as a useful therapeutic target in hepatoblastoma. Biosci Rep 2019;39:BSR20192466.
28Aronson DC, Czauderna P, Maibach R, Perilongo G, Morland B. The treatment of hepatoblastoma: Its evolution and the current status as per the SIOPEL trials. J Indian Assoc Pediatr Surg 2014;19:201-7.
29Huang J, Hu Y, Jiang H, Xu Y, Lu S, Sun F, et al. CHIC risk stratification system for predicting the survival of children with hepatoblastoma: Data from children with hepatoblastoma in China. Front Oncol 2020;10:552079.
30Zhi T, Zhang W, Zhang Y, Hu H, Huang D. Clinical characteristics and prognosis analysis of infantile hepatoblastoma – A 15-year retrospective single-center study. Cancer Manag Res 2021;13:3201-8.
31Waldherr M, Mišík M, Ferk F, Tomc J, Žegura B, Filipič M, et al. Use of HuH6 and other human-derived hepatoma lines for the detection of genotoxins: A new hope for laboratory animals? Arch Toxicol 2018;92:921-34.
32Huge N, Sandbothe M, Schröder AK, Stalke A, Eilers M, Schäffer V, et al. Wnt status-dependent oncogenic role of BCL9 and BCL9L in hepatocellular carcinoma. Hepatol Int 2020;14:373-84.
33Kim DH, Kim WD, Kim SK, Moon DH, Lee SJ. TGF-β1-mediated repression of SLC7A11 drives vulnerability to GPX4 inhibition in hepatocellular carcinoma cells. Cell Death Dis 2020;11:406.
34Clevers H, Nusse R. Wnt/β-catenin signaling and disease. Cell 2012;149:1192-205.
35Nusse R, Clevers H. Wnt/β-catenin signaling, disease, and emerging therapeutic modalities. Cell 2017;169:985-99.
36Zhang Y, Wang X. Targeting the Wnt/β-catenin signaling pathway in cancer. J Hematol Oncol 2020;13:165.
37Singh V, Singh AP, Sharma I, Singh LC, Sharma J, Borthakar BB, et al. Epigenetic deregulations of Wnt/β-catenin and transforming growth factor beta-Smad pathways in esophageal cancer: Outcome of DNA methylation. J Cancer Res Ther 2019;15:192-203.
38Yang Y, Zhou H, Zhang G, Xue X. Targeting the canonical Wnt/β-catenin pathway in cancer radioresistance: Updates on the molecular mechanisms. J Cancer Res Ther 2019;15:272-7.
39Li XY, Wen JY, Jia CC, Wang TT, Li X, Dong M, et al. MicroRNA-34a-5p enhances sensitivity to chemotherapy by targeting AXL in hepatocellular carcinoma MHCC-97L cells. Oncol Lett 2015;10:2691-8.
40Ma Y, Huang YX, Chen YY. miRNA34a5p downregulation of VEGFA in endometrial stem cells contributes to the pathogenesis of endometriosis. Mol Med Rep 2017;16:8259-64.
41Guo L, Zhong D, Lau S, Liu X, Dong XY, Sun X, et al. Sox7 is an independent checkpoint for beta-catenin function in prostate and colon epithelial cells. Mol Cancer Res 2008;6:1421-30.